摘要
运用非同位素银染mRNA差异显示方法 ,分析红莲型杂交水稻组合基因表达差异。以杂交水稻及其亲本三系 (YTA、YTB、93 1 1 )三叶一心期叶片RNA为模板采用锚定引物 5′dT1 2MN 3′和随机引物组合 ,进行反转录差异显示PCR。经 5 .6%测序胶电泳分离后用银染方法显示差异DNA条带 ,在直观下 ,从凝胶中回收差异条带并进行再扩增 ,2 %琼脂糖电泳得到了DNA的单一带型。
mRNA differential display polymerase chain reaction (DD-PCR)method with silver-staining, was used to isolate the differentially expressed gene of HL-type. Total RNA was extracted from 4 rice leaves of hybrid rice Honglianyou-6 and its parental three lines (Yuetai A、Yuetai B、9311)at the three-leaf stage.The cDNA copies of differentially expressed mRNA were identified by means of reverse transcription with anchor primer and amplified by subsequent PCR with sets of anchor primer and arbitrary primer.The DNA bands on gel were displayed by silver stain method after electrophoring in urea-denaturing sequencing gel.The differentially expressed bands were visually retrieved and reamplified , most of which obtained one band when visualized in 2% agarose gels.
出处
《江西农业大学学报》
CAS
CSCD
2003年第4期493-497,共5页
Acta Agriculturae Universitatis Jiangxiensis
基金
国家 973资助项目 ( 2 0 0 1CB1 0 880 6)
